JP6177249B2 - Compositions and methods for increasing muscle mass and strength by specifically antagonizing GDF8 and / or activin A - Google Patents

Description

  The present invention relates to compositions and methods for increasing the muscle mass and strength of a subject. More specifically, the invention relates to compositions that specifically bind GDF8 and activin A, and the use of such compositions to treat diseases and disorders characterized by muscle mass or muscle weakness.

  Growth differentiation factor 8 (GDF8, also called myostatin) is a secreted ligand belonging to the transforming growth factor β (TGF-β) superfamily of growth factors. GDF8 plays a central role in the development and maintenance of skeletal muscle that acts as a negative regulator of muscle mass. While the myostatin null mouse phenotype shows the importance of GDF8 in controlling mouse size at the developmental stage, muscle hypertrophy can also be induced in adult mice through inhibition of GDF8 by neutralizing antibodies, decoy receptors, or antagonists Can do. Administration of GDF8 neutralizing antibody has been reported to result in an increase in muscle mass between 10 and 30%. Increased muscle mass is seen due to increased fiber diameter as opposed to muscle fiber hyperplasia (number of fibers). Some studies have also reported an increase in muscle strength or performance commensurate with an increase in size, including spasm and twitch. Use of a cleavage resistant version of the GDF8 propeptide also results in an increase in muscle size.

  Other GDF8 antagonists are used in adult mice and have a significant effect on skeletal muscle mass. These include the extracellular portion of the type II GDF8 receptor, ActRIIB, stabilized by fusion to an IgG Fc domain (“ActRIIB-Fc”). The clinical molecule “ACE-031” is an example of an ActRIIB-Fc molecule.

  ActRIIB-Fc has been shown to increase muscle mass in experimental animals, but in human clinical trials this molecule has been shown to cause a variety of adverse side effects. For example, administration of ACE-031 to postmenopausal women in a phase Ib escalating dose study has been shown to result in an undesirable increase in hemoglobin and a decrease in FSH levels. Also, the phase II trial of ACE-031 in pediatric patients with muscular dystrophy was discontinued due to side effects including nasal and gingival bleeding. Vasodilation is also observed in patients treated with ActRIIB-Fc.

  From experiments, ActRIIB-Fc attenuates the muscle growth-inducing effect, but is not eliminated in myostatin null mice, so ActRIIB-Fc has its ability to antagonize one or more other ActRIIB ligands in addition to GDF8. This suggests that it has a muscle mass-inducing effect. Other ligands that bind ActRIIB include activin A, activin B, activin AB, inhibin A, inhibin B, GDF3, GDF11, Nodal, BMP2, BMP4, and BMP7.

  We hypothesized that ActRIIB-Fc enhanced muscle growth effects, as well as its undesirable side effects, were due to the binding of this molecule to additional ligands other than GDF8. Therefore, the inventors specifically antagonized only certain ActRIIB ligands in order to provide enhanced muscle growth effects of ActRIIB-Fc, while at the same time avoiding undesirable adverse side effects associated with this molecule, But we tried to determine if it was possible not to antagonize with others. Through experiments set out in the examples herein, it has been surprisingly discovered that significant muscle growth enhancement can be achieved by antagonizing specifically activin A. Importantly, the desired therapeutic effect of ActRIIB-Fc (eg, enhanced skeletal muscle growth) specifically antagonizes GDF8 and activin A, but other ActRIIB ligands (eg, GDF11, BMP9, BMP10, etc.) It has also been found that by not antagonizing the can be realized without undesirable side effects.

  Thus, according to one aspect of the invention, a composition comprising a GDF8 specific binding protein and an activin A specific binding protein is provided. In certain embodiments, the GDF8 specific binding protein is an anti-GDF8 antibody and / or the activin A specific binding protein is an anti-activin A antibody. According to a related aspect of the invention, there is provided an antigen binding molecule comprising a GDF8 specific binding domain and an activin A specific binding domain. In one embodiment of this aspect of the invention, the antigen binding molecule comprises a bispecific antibody comprising a first variable domain that specifically binds GDF8 and a second variable domain that specifically binds activin A. It is.

  The present invention provides a method of increasing a subject's muscle mass or strength by administering an activin A specific binding protein to the subject. The present invention also includes administering a GDF8-specific binding protein and an activin A-specific binding protein to a subject, or administering an antigen-binding molecule comprising a GDF8-specific binding domain and an activin A-specific binding domain to a subject. A method for increasing the muscle mass or strength of a subject is provided. The method according to this aspect of the invention is useful for treating diseases or disorders associated with muscle mass, muscle strength or muscle power loss, including, for example, cachexia, sarcopenia and other muscle wasting conditions.

  Other embodiments of the invention may become apparent from consideration of the following detailed description.

  Before describing the present invention, it will be appreciated that such methods and conditions may vary, and the invention is not limited to the specific methods and experimental conditions described. It will also be appreciated that the terminology used herein is for the purpose of describing particular embodiments only, and since the scope of the invention can be limited only by the scope of the appended claims. It is not intended to be limiting.

  Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. As used herein, the term “about” when used in connection with a particular recited numerical value means that the value may vary from the recited value by as little as 1%. For example, as used herein, the expression “about 100” means any value between 99 and 101 and (eg, 99.1, 99.2, 99.3, 99.4, etc.). Including.

TECHNICAL FIELD The present invention relates to a composition comprising an antigen-specific binding protein. More specifically, the present invention provides a composition comprising a GDF8 specific binding protein and an activin A specific binding protein.

  As used herein, the expression “antigen-specific binding protein” means a protein comprising at least one domain that specifically binds a particular antigen. Exemplary categories of antigen-specific binding proteins include antibodies, antigen-binding portions of antibodies, peptides that specifically interact with specific antigens (eg, peptibodies), receptor molecules that specifically interact with specific antigens, And a protein comprising a ligand binding portion of a receptor that specifically binds a particular antigen.

  The present invention includes antigen-specific binding proteins that specifically bind GDF8, ie, “GDF8-specific binding proteins”. The term “GDF8” (also called “growth differentiation factor 8” and “myostatin”) means a protein having the amino acid sequence of SEQ ID NO: 25 (mature protein). According to the present invention, GDF8-specific binding protein specifically binds GDF8, but other GDF3, BMP2, BMP4, BMP7, BMP9, BMP10, GDF11, Activin A, Activin B, Activin AB, Nodal, etc. Does not bind ActRIIB ligand.

  The present invention also includes antigen-specific binding proteins that specifically bind activin A, ie, “activin A-specific binding proteins”. Activins are homo and heterodimeric molecules that contain βA and / or βB subunits. The βA subunit has the amino acid sequence of SEQ ID NO: 26, and the βB subunit has the amino acid sequence of SEQ ID NO: 28. Activin A is a homodimer of two βA subunits; Activin B is a homodimer of two βB subunits; and Activin AB is a heterodimer of one βA subunit and one βB subunit. It is a mer. The activin A specific binding protein can be an antigen specific binding protein that specifically binds the βA subunit. Since the βA subunit is found in both activin A and activin AB molecules, an “activin A specific binding protein” can be an antigen-specific binding protein that specifically binds activin A and activin AB (βA Thanks to its interaction with subunits). Thus, according to the present invention, the activin A specific binding protein specifically binds activin A or activin A and activin AB, but activin B, GDF3, GDF8, BMP2, BMP4, BMP7, BMP9, BMP10, It does not bind other ActRIIB ligands such as GDF11, Nodal.

  In the context of the present invention, a molecule such as ActRIIB-Fc (eg, “ACE-031”) comprising a ligand binding portion of an ActRIIB receptor is a plurality of molecules other than GDF8, Activin A and Activin AB. Because it binds a ligand, it is not considered a “GDF8-specific binding protein” or “activin A-specific binding protein”.

Antigen-binding molecules with two different antigen-specific binding domains The invention also includes antigen-binding molecules that comprise two different antigen-specific binding domains. In particular, the present invention includes antigen binding molecules comprising a GDF8 specific binding domain and an activin A specific binding domain. As used herein, the term “antigen-specific binding domain” refers to (i) an antigen-binding fragment of an antibody molecule, (ii) a peptide (eg, a peptibody) that specifically interacts with a particular antigen, and / or (Iii) a polypeptide comprising or consisting of: a ligand binding portion of a receptor that specifically binds a particular antigen. For example, the present invention provides one arm comprising a first heavy chain variable region / light chain variable region (HCVR / LCVR) pair that specifically binds GDF8, and a second HCVR that specifically binds activin A. Bispecific antibodies with separate arms containing / LCVR pairs.

Specific binding As used herein, the term “specifically binds” and the like refers to an antigen-specific binding protein, or a specific binding domain whose antigen-specific binding domain is characterized by a dissociation constant (K _D ) of 500 pM or less. It means forming a complex with the antigen and not binding other unrelated antigens under normal test conditions. An “irrelevant antigen” is a protein, peptide or polypeptide that has less than 95% amino acid identity to each other. Methods for determining whether two molecules specifically bind to each other are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. For example, an antigen-specific binding protein or antigen-specific binding domain used in the context of the present invention is less than about 500 pM, less than about 400 pM, less than about 300 pM, about 200 pM, as measured by a surface plasmon resonance assay. Less, less than about 100 pM, less than about 80 pM, less than about 70 pM, less than about 60 pM, less than about 50 pM, less than about 30 pM, less than about 20 pM, less than about 10 pM, less than about 5 pM, about 4 pM less than about 2pM less than about 1pM less than about 0.5pM less than about 0.2pM less than about 0.1 pM, or about 0.05pM less than the specific antigen with a K _D (e.g. , GDF8, or activin A and / or AB) or parts thereof Including the molecule.

As used herein, antigen-specific binding protein or antigen-specific binding domains, when tested for binding to molecules at 25 ° C. in the surface plasmon resonance assay, proteins or binding domains, 1000 pM greater than K _D Or cannot show any binding in such an assay or equivalent thereof, “does not bind” to a particular molecule (eg, “does not bind GDF11”, “does not bind BMP9”, “BMP10”). Do not combine ").

  The term “surface plasmon resonance” as used herein refers to the detection of changes in protein concentration within a biosensor matrix, for example using the BIAcore ™ system (Biacore Life Sciences division of GE Healthcare, Piscataway, NJ). An optical phenomenon that enables real-time interaction analysis.

The term “K _D ” as used herein refers to the equilibrium dissociation constant of a particular protein-protein interaction (eg, antibody-antibody interaction). Unless indicated otherwise, K _D values disclosed herein, refers to a K _D values measured by surface plasmon resonance assay at 25 ° C..

Antibodies and Antigen-binding Fragments of Antibodies As indicated above, antigen-specific binding proteins can comprise or consist of antibodies or antigen-binding fragments of antibodies. Furthermore, in the case of antigen binding molecules comprising two different antigen-specific binding domains, one or both antigen-specific binding domains may comprise or consist of an antigen-binding fragment of an antibody.

The term “antibody” as used herein refers to four polypeptide chains, two heavy (H) chains and two light (L) chains linked by disulfide bonds, and multimers thereof (eg, IgM). Is intended to refer to an immunoglobulin molecule comprising Each heavy chain includes a heavy chain variable region (abbreviated herein as HCVR or V _H ) and a heavy chain constant region. The heavy chain constant region includes three domains, C _H 1, C _H 2 and C _H 3. Each light chain includes a light chain variable region (abbreviated herein as LCVR or _VL ) and a light chain constant region. The light chain constant region contains one domain (C _L 1). The V _H and V _L regions can be further subdivided into hypervariable regions called complementarity determining regions (CDRs) and can be interspersed with more conserved regions called framework regions (FR). Each V _H and V _L region is composed of three CDRs and four FRs, arranged in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 from the amino terminus to the carboxy terminus. In different embodiments of the invention, the FR (or antigen-binding portion thereof) of an antibody of the invention can be identical to a human germline sequence or can be naturally or artificially modified. Amino acid consensus sequences can be defined based on a parallel analysis of two or more CDRs.

  As used herein, the term “antibody” also includes antigen-binding fragments of complete antibody molecules. As used herein, the terms “antigen-binding portion” of an antibody, “antigen-binding fragment” of an antibody, and the like refer to naturally occurring, enzymatically that specifically binds an antigen to form a complex. Any polypeptide, glycoprotein, available, synthetic or genetically modified is included. An antigen-binding fragment of an antibody can be obtained using any suitable standard technique, such as proteolysis, or recombinant genetic engineering techniques that involve manipulation and expression of DNA encoding antibody variable and optionally constant domains. It can be derived from a molecule. Such DNA is known and / or readily available from, for example, commercial sources, DNA libraries (eg, including phage-antibody libraries), or can be synthesized. DNA to place one or more variable and / or constant domains in the proper configuration, or to introduce codons, create cysteine residues, modify amino acids, etc., add or delete Can be sequenced and manipulated chemically or by using molecular biology techniques.

  Non-limiting examples of antigen binding fragments are: (i) Fab fragment; (ii) F (ab ′) 2 fragment; (iii) Fd fragment; (iv) Fv fragment; (v) single chain Fv (scFv) molecule; vi) dAb fragments; and (vii) minimal recognition consisting of hypervariable regions of antibodies (eg, isolated complementarity determining regions (CDRs) such as CDR3 peptides), or amino acid residues that mimic constrained FR3-CDR3-FR4 peptides Includes: Domain-specific antibody, single domain antibody, domain-deficient antibody, chimeric antibody, CDR-grafted antibody, bispecific antibody, trispecific antibody, tetraspecific antibody, minibody, nanobody (eg monovalent nanobody, two Other engineered molecules such as valency Nanobodies), small modular immunopharmaceuticals (SMIPs), and shark variable IgNAR domains are also encompassed within the expression “antigen-binding fragment” as used herein.

An antigen-binding fragment of an antibody will usually contain at least one variable domain. The variable domain may be of any size and amino acid composition and generally comprises at least one CDR adjacent to or within the frame with one or more framework sequences. Let's go. In antigen-binding fragments that have a V _H domain associated with a V _L domain, the V _H and V _L domains can be located in any suitable sequence relative to each other. For example, the variable region is a dimer and may contain a _VH - _VH , _VH - _VL or _VL - _VL dimer. Alternatively, an antigen-binding fragment of an antibody can contain monomeric V _H and V _L domains.

In certain embodiments, an antigen-binding fragment of an antibody can contain at least one variable domain covalently linked to at least one constant domain. Non-limiting exemplary configurations of variable and constant domains that can be found within antigen-binding fragments of the antibodies of the invention are: (i) V _H -C _H 1; (ii) V _H -C _H 2; _{iii) V H -C H 3;} (iv) V H -C H 1-C H 2; (v) V H -C H 1-C H 2-C H 3; (vi) V H -C H 2 _{-C H 3; (vii) V} H -C L; (viii) V L -C H 1; (ix) V L -C H 2; (x) V L -C H 3; (xi) V L - _{C H 1-C H 2;} (xii) V L -C H 1-C H 2-C H 3; (xiii) V L -C H 2-C H 3; and (xiv) V _L -C _L: including. In any configuration of variable and constant domains, including any of the exemplary configurations listed above, the variable and constant domains can be directly linked to each other or linked by a complete or partial hinge or linker region. obtain. The hinge region is at least two (e.g., 5, 10, 15, 20, 40, 60 or) that provide a flexible or semi-flexible link between adjacent variable and / or constant domains in a single polypeptide molecule. It can consist of amino acids). Moreover, antigen-binding fragments of the antibodies of the invention can be non-covalently associated with each other and / or with one or more monomeric V _H or V _L domains (eg, one or more disulfides). Binding), homodimers or heterodimers (or other multimers) of any of the variable and constant domain configurations listed above.

  The molecules of the invention may comprise or consist of human antibodies and / or recombinant human antibodies, or fragments thereof. The term “human antibody” as used herein includes antibodies having variable and constant regions derived from human germline immunoglobulin sequences. Human antibodies are still encoded by human germline immunoglobulin sequences (eg mutations introduced by random or site-directed mutagenesis in vitro or by somatic mutation in vivo), eg in CDR and in particular in CDR3. Amino acid residues that are not converted may be included. However, as used herein, the term [human antibody] is intended to include antibodies in which the germline-derived CDR sequences of another mammalian species such as mouse are grafted onto the human framework sequences. It is not a thing.

The molecule of the invention may comprise or consist of a recombinant human antibody or antigen-binding fragment thereof. The term “recombinant human antibody” as used herein refers to an antibody expressed using a recombinant expression vector transfected into a host cell (as described further below), a recombinant combinatorial human antibody library (further Antibodies, isolated from human immunoglobulin genes (see eg Taylor et al. (1992) Nucl. Acids Res. 20: 6287-6295) isolated from animals (eg mice) isolated from Prepared by recombinant means, such as an antibody to be isolated, or an antibody prepared, expressed, created or isolated by other means, including splicing of human germline immunoglobulin gene sequences to other DNA sequences It is intended to include all human antibodies that are expressed, made or isolated. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. In certain embodiments, however, such recombinant human antibodies have been subjected to in vitro mutagenesis (or in vivo somatic mutagenesis when transgenic animals to human Ig sequences are used) and are therefore recombinant antibodies. The amino acid sequences of the V _H and V _L regions of this antibody may be derived from and related to the human germline V _H and V _L sequences, while they may not exist naturally within the human antibody germline repertoire in vivo. is there.

Anti-GDF8 Antibodies and Antigen Binding Fragments thereof In certain embodiments of the invention, the GDF8 specific binding protein, or GDF8 specific binding domain comprises or consists of an anti-GDF8 antibody or antigen binding fragment thereof. Anti-GDF8 antibodies are, for example, US Pat. Nos. 6,096,506; 7,320,789; 7,261,893; 7,807,159; 7,888,486; 7,635,760; 499; US Patent Application Publication Nos. 2007/0178095; 2010/0166764; 2009/0148436; and International Patent Application Publication No. WO 2010/070094. Anti-GDF8 antibodies are also described in US Patent Application No. 13 / 115,170, filed May 25, 2011 and published as US2011 / xxxxxxxx, and named 8D12, H4H1657N2, and H4H1669P. Contains antibodies. Any of the anti-GDF8 antibodies, or antigen-binding fragments thereof, mentioned or described in any of the foregoing patents or publications, such antibodies and / or antigen-binding fragments “specifically bind” GDF8. Insofar as its expression can be used in the context of the present invention, as defined herein.

  Table 1 shows HCVR, LCVR, and CDR sequence identifiers of certain non-limiting exemplary anti-GDF8 antibodies that can be used in the context of the present invention.

Anti-activin A antibody and antigen-binding fragment thereof In a specific embodiment of the present invention, activin A-specific binding protein or activin A-specific binding domain is an antibody or antigen-binding fragment thereof that specifically binds activin A. Comprising or consisting of. In certain embodiments, the activin A specific binding protein specifically binds the βA subunit. An antigen-specific binding protein that specifically binds the βA subunit can recognize both activin A (βA / βA homodimer) and activin AB (βA / βB heterodimer). Thus, according to the present invention, the activin A specific binding protein can bind either activin A or activin AB (not activin B). Anti-activin A antibodies are mentioned, for example, in US Patent Application Publication No. 2009/0234106. A specific anti-activin A antibody is designated “MAB3381” and is commercially available from R & D Systems, Inc, Minneapolis, Minn. MAB3381 specifically binds activin A (homodimer) as well as activin AB (heterodimer). Any of the aforementioned anti-activin A antibodies, or antigen-binding fragments thereof, can be a “specific” activin A and / or activin AB, as such antibodies and / or antigen-binding fragments are defined herein. Can be used in the context of the present invention.

Pharmaceutical compositions and methods of administration The present invention includes pharmaceutical compositions comprising a GDF8 specific binding protein and an activin A specific binding protein. The invention also includes a pharmaceutical composition comprising an antigen binding molecule comprising a GDF8 specific binding domain and an activin A specific binding domain. The pharmaceutical compositions of the invention are formulated with suitable carriers, excipients, and other agents that provide suitable transport, delivery, tolerance, and the like. A number of suitable formulations can be found, for example, in Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton PA. Suitable formulations include, for example, powders, pastes, ointments, jellies, waxes, oils, lipids, lipids (cations or anions) containing vesicles (such as LIPOFECTIN ™), DNA complexes, anhydrous absorbent pastes, water Oil-type and water-in-oil emulsions, emulsion carbowaxes (polyethylene glycols of various molecular weights), semisolid gels, and semisolid mixtures containing carbowax. Further suitable formulations are also described in Powell et al. “Compendium of containers for parenteral formulations” PDA (1998) J Pharm Sci Technol 52: 238-311.

  Various delivery systems such as liposome encapsulation, microparticles, microcapsules, recombinant cells capable of expressing mutant viruses, receptor-mediated endocytosis (eg, Wu et al., 1987, J. Biol Chem. 262: 4429-4432) are known and can be used to administer the pharmaceutical compositions of the present invention. Methods of administration include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The composition may be administered by any conventional route, for example by infusion and bolus injection, by absorption through epithelial and dermal mucosal linings (eg, oral mucosa, rectum and intestinal mucosa, etc.) and other It may be administered with a bioactive agent.

  The pharmaceutical compositions of the invention can be delivered subcutaneously or intravenously using standard needles and syringes. Also, for subcutaneous delivery, pen delivery devices have an easy use for delivering the pharmaceutical compositions of the present invention. Such a pen delivery device may be reusable or disposable. Reusable pen delivery devices generally utilize replaceable cartridges that contain a pharmaceutical composition. Once all of the pharmaceutical composition in the cartridge has been dispensed and the cartridge is empty, the empty cartridge can be easily discarded and replaced with a new cartridge containing the pharmaceutical composition. The pen delivery device can then be reused. In disposable pen delivery devices, there are no replaceable cartridges. Rather, the disposable pen delivery device becomes prefilled with a pharmaceutical composition held in a reservoir within the device. Once the reservoir pharmaceutical composition is empty, the entire device is discarded.

  A number of reusable pen-type and self-injector delivery devices have application for subcutaneous delivery of the pharmaceutical compositions of the present invention. Examples include, but are not limited to, AUTOPEN ™ (Owen Mumford, Inc., Woodstock, UK), DISETRONIC ™ pen (Disetronic Medical Systems, Bergdorf, Switzerland) to name a few. ), HUMALOG MIX 75/25 (TM) pen, HUMALOG (TM) pen, HUMALIN 70/30 (TM) pen (Eli Lilly and Co., Indianapolis, IN), NOVOPEN (TM) I, II and III (Novo Nordisk , Copenhagen, Denmark), NOVOPEN JUNIOR (TM) (Novo Nordisk, Copenhagen, Denmark), BD (TM) pen (Becton Dickinson, Franklin Lakes, NJ), OPTIPEN (TM), OPTIPEN PRO (TM), OPTIPEN STARLET (TM) And OPTICLIK ™ (sanofi-aventis, Frankfurt, Germany). Examples of disposable pen delivery devices that have use in the subcutaneous delivery of pharmaceutical compositions of the present invention include, but are not limited to, SOLOSTAR ™ pen (sanofi-aventis), to name a few: FLEXPEN ™ (Novo Nordisk), and KWIKPEN ™ (Eli Lilly), SURECLICKTM Autoinjector (Amgen, Thousand Oaks, CA), PENLETTM ™ (Haselmeier, Stuttgart, Germany), EPIPEN (Dey, LP), and HUMIRATM pen (Abbott Labs, Abbott Park IL).

  In certain situations, the pharmaceutical composition of the invention can be delivered in a controlled release system. In one embodiment, a pump may be used (see Langer, supra; Sefton, 1987, CRC Crit. Ref. Biomed. Eng. 14: 201). In another embodiment, polymeric materials may be used; see Medical Applications of Controlled Release, Langer and Wise (eds.), 1974, CRC Pres., Boca Raton, Florida. In yet another embodiment, the controlled release system can be placed in close proximity to the target of the composition and therefore requires only a fraction of the systemic dose (eg, Goodson, 1984, in Medical Applications of Controlled Release supra, vol. 2, pp. 115-138). Other controlled release systems are discussed in a review by Langer, 1990, Science 249: 1527-1533.

  Injectable formulations may include dosage forms for intravenous, subcutaneous, intradermal and intramuscular injections, drops, and the like. These injectable preparations can be prepared by known methods. For example, an injectable preparation can be prepared, for example, by dissolving, suspending or emulsifying the above-described antibody or a salt thereof in a sterile aqueous medium or oily medium usually used for injection. Examples of the aqueous medium for injection include alcohol (for example, ethanol), polyhydric alcohol (for example, propylene glycol, polyethylene glycol), nonionic surfactant [for example, polysorbate 80, HCO-50 (polyoxygen of hydrogenated castor oil) For example, isotonic solutions containing saline, glucose or other adjuvants, which may be used in combination with a suitable solubilizer such as ethylene (50 mol adduct). Examples of the oil medium include sesame oil and soybean oil which can be used in combination with a solubilizer such as benzyl benzoate and benzyl alcohol. The injection prepared in this way is preferably filled in a suitable ampoule.

  Advantageously, the above-mentioned oral or parenteral pharmaceutical composition is formulated in a unit dosage form suitable to suit the dose of the active ingredient. Such unit dose dosage forms include, for example, tablets, pills, capsules, injections (ampoules), suppositories and the like.

Dosage The amount of active ingredient (eg, anti-GDF8 antibody and anti-activin A antibody) that can be administered to a subject is generally a therapeutically effective amount. As used herein, the phrase “therapeutically effective amount” refers to one or more of the following parameters: weight, muscle mass (eg, anterior tibialis [TA] muscle mass, gastrocnemius [GA] muscle Means the dose of antigen-specific binding protein and / or antigen-binding molecule that results in a detectable increase in volume, quadriceps [Quad] muscle mass, etc., muscle strength / muscle power, and / or muscle function. For example, a “therapeutically effective amount” of a GDF8-specific binding protein and / or activin A-specific binding protein is administered to a test subject when treated with a control, eg, as shown in Example 1 herein. Results in a TA or GA muscle mass increase of at least 2%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60% or more compared to the treated subject, For example, it includes the amount of GDF8 specific binding protein and / or activin A specific binding protein.

  In the case of an antibody of the invention (eg, an anti-GDF8 antibody, an anti-activin A antibody, or a bispecific antibody that specifically binds GDF8 and activin A), a therapeutically effective amount is about 0. 05 mg to about 600 mg; for example, about 0.05 mg, about 0.1 mg, about 1.0 mg, about 1.5 mg, about 2.0 mg, about 10 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, About 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 210 mg, about 220 mg, about 230 mg About 240 mg, about 250 mg, about 260 mg, about 270 mg, about 280 mg, about 290 mg, about 00 mg, about 310 mg, about 320 mg, about 330 mg, about 340 mg, about 350 mg, about 360 mg, about 370 mg, about 380 mg, about 390 mg, about 400 mg, about 410 mg, about 420 mg, about 430 mg, about 440 mg, about 450 mg, about 460 mg, It may be about 470 mg, about 480 mg, about 490 mg, about 500 mg, about 510 mg, about 520 mg, about 530 mg, about 540 mg, about 550 mg, about 560 mg, about 570 mg, about 580 mg, about 590 mg, or about 600 mg.

  The amount of antibody of the invention (eg, anti-GDF8 antibody, anti-activin A antibody, or bispecific antibody that specifically binds GDF8 and activin A) contained within an individual dose is per kilogram of patient body weight. The antibody may be expressed in milligrams (ie mg / kg). For example, an anti-GDF8, anti-activin A and / or anti-GDF8 / anti-activin A bispecific antibody of the present invention may have a dose of about 0.0001 to about 50 mg / kg of patient weight (eg, 0.5 mg / kg, 4. 1.0 mg / kg, 1.5 mg / kg, 2.0 mg / kg, 2.5 mg / kg, 3.0 mg / kg, 3.5 mg / kg, 4.0 mg / kg, 4.5 mg / kg 0 mg / kg, 5.5 mg / kg, 6.0 mg / kg, 6.5 mg / kg, 7.0 mg / kg, 7.5 mg / kg, 8.0 mg / kg, 8.5 mg / kg, 9.0 mg / kg, 9.5 mg / kg, 10.0 mg / kg, 10.5 mg / kg, 11.0 mg / kg, 11.5 mg / kg, etc.).

  The composition of the invention may comprise equal amounts of GDF8-specific binding protein and activin A-specific binding protein. Alternatively, the amount of GDF8 specific binding protein in the composition may be less than or greater than the amount of activin A specific binding protein. One skilled in the art can use routine experimentation to determine the appropriate amount of each individual component in the composition of the invention necessary to provide the desired therapeutic effect.

Therapeutic methods The present invention relates to increasing individual muscle strength / muscle power and / or muscle mass and / or muscle function or by specifically binding GDF8 and / or activin A and / or activin AB. And methods of treating conditions or afflictions that can be cured, reduced or ameliorated by beneficially altering metabolism (carbohydrate, lipid and protein processing) without binding other ActRIIB ligands. For example, the present invention is characterized by increasing a subject's muscle strength / muscle power and / or muscle mass and / or muscle function, or reducing a subject's muscle mass or muscle strength by administering an activin A-specific binding protein to the subject. A method of treating a disease or disorder. The present invention also increases the muscle strength / muscle power and / or muscle mass and / or muscle function of a subject, or muscle mass of a subject by administering a GDF8-specific binding protein and an activin A-specific binding protein to the subject. Or a method of treating a disease or disorder characterized by muscle weakness. Any of the GDF8 specific binding proteins and / or activin A specific binding proteins disclosed or described herein can be used in the context of these aspects of the invention. For example, the treatment method of the present invention includes administering an anti-GDF8 antibody and / or an anti-activin A antibody to a subject.

  In a method comprising administering a GDF8-specific binding protein and an activin A-specific binding protein to a subject, the GDF8-specific binding protein and the activin A-specific binding protein are at the same or substantially the same time to the subject, eg, simply It can be administered in multiple therapeutic doses or in two separate doses that are administered simultaneously or within less than about 5 minutes of each other. Alternatively, the GDF8-specific binding protein and activin A-specific binding protein can be administered to the subject sequentially, eg, at separate medical dosages that are separated from each other for a time longer than about 5 minutes.

  The present invention also increases the muscle strength / muscle power and / or muscle mass and / or muscle function of a subject by administering to the subject an antigen binding molecule comprising a GDF8 specific binding domain and an activin A specific binding domain. Or a method of treating a disease or disorder characterized by decreased muscle mass or weakness in a subject. Any of the antigen binding molecules disclosed or referred to herein can be used in the context of this aspect of the invention. For example, the therapeutic method of the present invention comprises a first variable domain comprising an HCVR / LCVR pair that specifically binds GDF8 and a second variable domain comprising an HCVR / LCVR pair that specifically binds activin A. Administering a bispecific antibody to the subject.

  The composition of the present invention comprises one or more additional therapeutic agents including, for example, growth factor inhibitors, immunosuppressants, anti-inflammatory agents, metabolic inhibitors, enzyme inhibitors, cytotoxic / cytostatic agents Can be administered to a subject. The additional therapeutic agent or agents can be administered prior to, simultaneously with, or after administration of the GDF8 and activin A specific binding proteins of the invention.

  Exemplary diseases, disorders and conditions that can be treated with the compositions of the present invention include, but are not limited to, sarcopenia, cachexia (other conditions such as cancer, chronic renal failure, or chronic obstructive Idiopathic or secondary to lung disease), muscle damage, muscle wasting and atrophy, eg disuse, immobilization, bed rest, injury, medical treatment or surgical intervention (eg hip fracture, hip prosthesis) Including muscle atrophy or muscle wasting caused or associated with the need for replacement, knee replacement, etc.) or artificial respiration. Compositions of the invention also include cancer, obesity, diabetes, arthritis, multiple sclerosis, muscular dystrophy, amyotrophic lateral sclerosis, Parkinson's disease, osteoporosis, osteoarthritis, metabolic syndrome (but not limited to Such as obesity, malnutrition, organ atrophy, chronic obstructive pulmonary disease, and anorexia) can be used to treat, prevent or ameliorate.

Avoiding side effects The present invention relates to a subject's strength / muscle power and / or muscle mass and / or mass without causing adverse side effects associated with administration of molecules that bind multiple (eg, 3 or more) ActRIIB ligands. And / or methods of treating a disease or disorder characterized by increased muscle function or reduced muscle mass or weakness in a subject. For example, a clinical molecule called ACE-031 (Acceleron Pharma, Inc., Cambridge, MA) consists of the extracellular portion of ActRIIB fused to an IgG Fc domain (also referred to herein as “ActRIIB-Fc”). It is a multimer. ActRIIB-Fc binds GDF8 and other ActRIIB ligands such as, for example, activin A, activin B, GDF11, BMP9, BMP10, and TGFβ, and is known to cause various adverse side effects when administered to human patients It has been. Significantly, we specifically inhibited GDF8 and activin A (eg, by administering anti-GDF8 and anti-activin A antibodies), while activin B, GDF11, BMP9. , BMP10, and other ActRIIB ligands, such as TGFβ, at least equivalent to that observed by administration of ActRIIB-Fc without causing adverse side effects associated with non-specific activin binding agents such as ActRIIB-Fc Unexpectedly found to increase muscle mass.

Administration regimen According to one embodiment of the invention, a composition of the invention (eg comprising GDF8 and / or activin A specific binding protein or antigen binding molecule comprising a GDF8 specific binding domain and an activin A specific binding domain). Multiple administrations of the composition) can be administered to a subject over a defined time course. The method according to this aspect of the invention comprises sequentially administering to the subject multiple doses of the composition of the invention. As used herein, the term “sequentially administered” means that each dose of the composition of the invention is separated into the subject at different time points, eg, at predetermined intervals (eg, hours, days, weeks or months). Means administered on different days. The present invention includes the sequential administration of an initial dose of a composition of the invention to a patient, followed by one or more secondary doses of the composition, and optionally followed by one or more of the compositions. A method comprising administering a tertiary dose of

  The terms “initial dose”, “secondary dose”, and “tertiary dose” refer to the time sequence of administration of the composition of the present invention. Thus, the “initial dose” is the dose administered at the beginning of the treatment plan (also referred to as the “baseline dose”); the “secondary dose” is the dose administered after the first dose; and the “third dose” is The dose administered after the second dose. Initial, secondary and tertiary doses may all contain the same amount of one or more active ingredients but will generally differ from one another with respect to dosing frequency. However, in certain embodiments, the amount of the active ingredient or ingredients contained in the initial, secondary and / or tertiary doses will differ from one another during the course of treatment (eg, above or Adjusted below).

  In one exemplary embodiment of the invention, each secondary and / or tertiary dose is 1-30 (eg 1, 2, 3, 4, 5, 6, 7, 8, 9, (10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or more) The As used herein, the phrase “immediate dose” refers to a composition of the invention that is administered to a subject in a series of multiple doses, in the order of no intervention dose, just prior to the next dose. One or more doses are meant.

  The method according to this aspect of the invention may comprise administering to the patient any number of secondary and / or tertiary doses of the composition of the invention. For example, in certain embodiments, only a single secondary dose is administered to a patient. In other embodiments, two or more (eg, 2, 3, 4, 5, 6, 7, 8, or more) secondary doses are administered to the patient. Similarly, in certain embodiments, only a single third dose is administered to a patient. In other embodiments, two or more (eg, 2, 3, 4, 5, 6, 7, 8, or more) tertiary doses are administered to the patient.

  In embodiments comprising multiple secondary doses, each secondary dose can be administered as often as the other secondary doses. For example, each secondary dose can be administered to the patient 1-29 days after the previous dose. Similarly, in embodiments involving multiple tertiary doses, each tertiary dose can be administered as often as the other tertiary doses. For example, each tertiary dose can be administered to the patient 1-60 days after the immediately preceding dose. Alternatively, the frequency with which secondary and / or tertiary doses are administered to a patient can vary over the course of the treatment plan. The frequency of administration can also be adjusted during the course of treatment by the physician according to individual patient requirements after clinical examination.

  The following examples are presented to provide one of ordinary skill in the art with a complete disclosure and description of how to make and use the methods and compositions of the present invention, and to the extent that the inventors regard as those inventions. It is not intended to be limiting. Efforts have been made to ensure accuracy with respect to numbers used (eg amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Centigrade, and pressure is at or near atmospheric.

Example 1 Specific inhibition of GDF8 and activin A results in a synergistic increase in skeletal muscle mass Introduction ActRIIB-Fc is a GDF8 antagonist consisting of the extracellular portion of the ActRIIB receptor stabilized by fusion to the IgGFc domain . ActRIIB-Fc has been shown to increase mouse muscle mass to a greater extent than anti-GDF8 antibodies. We hypothesized that the enhanced activity of ActRIIB-Fc may be due to its ability to bind additional ActRIIB other than GDF8. In particular, we proposed that Activin A antagonism, in addition to GDF8 antagonism, may lead to a greater increase in skeletal muscle mass than that observed in animals treated with anti-GDF8 antibody alone. . Therefore, this example is designed to determine whether specific inhibition of GDF8 and activin A can increase skeletal muscle mass to some extent at least equivalent to the increase observed with ActRIIB-Fc. It was.

Results and Discussion The degree of skeletal muscle hypertrophy induced by the administration of ActRIIB-Fc was compared with the administration effect of a combination of GDF8 specific antibody, activin A specific antibody, or anti-GDF8 + antiactivin A antibody. The ActRIIB-Fc construct used in this example has the amino acid sequence of SEQ ID NO: 27. The anti-GDF8 antibody used in this example is an antibody designated H4H1657N2 (see Table 1). The anti-activin A antibody used in this example is an antibody designated MAB3381 (available from R & D Systems, Inc., Minneapolis, MN). An isotype matched (hIgG4) antibody was used as a negative control.

  Briefly, 25 male CB17 SCID mice approximately 10 weeks old were treated (isotype control mAb, ActRIIB-Fc, H4H1657N2, MAB3381 or H4H1657N2 + MAB3381 equally divided into 5 groups based on body weight). Reagents were administered subcutaneously at a dose of 10 mg / kg twice during the first week (on days 0 and 3) and during the next 3 weeks (on days 7, 14 and 21) One dose was administered. On day 28, mice were euthanized and weighed, and the anterior tibial (TA) and gastrocnemius (GA) muscles were dissected and weighed. Tissues were normalized to starting body weight and the percent change in weight across isotype matched (hIgG4) control antibodies was calculated. The results are summarized in Table 2 and expressed as a percentage increase over the negative control ± standard error of the mean.

  To confirm that muscle hypertrophy was the result of increased muscle fiber size, anterior tibial (TA) muscle was implanted in OCT and frozen with isopentane for histological examination and immunohistochemical labeling. TA muscle cross sections were stained with anti-laminin antibody to outline myofibers and mean cross sectional area (CSA) was determined by using an image analysis system. The results of two independent experiments (Exp # 1 and Exp # 2) are summarized in Table 2B. All data were expressed as mean ± standard error of the mean.

  As shown in Table 2A, ActRIIB-Fc induced significant hypertrophy in all muscles examined, a 44.88 ± 5.35% increase in TA muscle mass, and 34.25 ± in GA muscle mass. An increase of 6.97% was shown. Treatment with H4H1657N2 (anti-GDF8) or MAB3381 (anti-activin A) alone also resulted in TA muscle mass (22.42 ± 1.65% and 19.09 ± 2.04%, respectively) and GA muscle mass (respectively, 24.17 ± 1.84 and 14.02 ± 0.91%) induced significant hypertrophy but was not as prominent as ActRIIB-Fc. However, the combination of H4H1657N2 and MAB3381 increased TA (55.13 ± 5.16%) and GA (41.72 ± 3.63%) even greater than those observed in ActRIIB-Fc treated animals Induced. Furthermore, it was confirmed that the observed muscle hypertrophy was the result of an increase in muscle fiber size (see Table 2B).

  Importantly, the extent of weight, TA and GA muscle gains for the anti-GDF8 + anti-activin A combination is substantially greater than the sum of these parameter increases observed in anti-GDF8 + anti-activin A monotherapy subjects. It was big. Thus, combined inhibition of GDF8 and activin A resulted in a synergistic increase in body weight and skeletal muscle mass, and these increases were more pronounced than those observed in ActRIIB-Fc treated animals. Moreover, as shown in the examples below, the increase in body weight and skeletal muscle mass of animals treated with GDF8 and activin A specific binding agents causes adverse side effects observed with molecules such as ActRIIB-Fc. Can be achieved without any trouble.

Example 2 Specific Antagonism of GDF8 and Activin A Does Not Cause Deleterious Side Effects Associated with Nonspecific Activin Ligand Binding Agents ActRIIB-Fc binds multiple ActRIIBs and causes significant side effects. This example demonstrated that the deleterious side effects associated with ActRIIB-Fc can be avoided by selectively antagonizing only one ActRIIB ligand, namely GDF8 and / or Activin A. In particular, in vivo erythrocyte properties (ie, increased endoglin levels and increased erythrocyte distribution width) that appear to be related to biomarkers, protein expression studies, and ActRIIB-Fc side effects in humans are only found in animals treated with ActRIIB-Fc. Although not seen in animals treated with anti-GDF8 antibody, anti-activin A antibody or a combination of anti-GDF8 antibody and anti-activin A antibody. Thus, taken together, the following results show that the specific antagonism of GDF8 and / or Activin A does not antagonize other ActRIIB ligands such as Activin B, GDF11, BMP9, BMP10, and / or TGFβ. , Which did not result in the undesirable phenotype associated with ActRIIB-Fc.

Results and Discussion Example 1 (ie, at the dose of 10 mg / kg twice during the first week [on days 0 and 3] and during the next 3 weeks [7 days, 14 days and 21 days Hematology using mice treated with a combination of ActRIIB-Fc (SEQ ID NO: 27), H4H1657N2 (anti-GDF8), MAB3381 (anti-activin A), or H4H1657N2 + MAB3381 according to the dosing schedule described in Research was conducted. Specifically, hemoglobin levels and red blood cell distribution width (RDW) (an indicator of certain blood disorders such as anemia) were measured from blood samples taken from mice treated with various drugs. RDW results (normalized to isotype control values) are summarized in Table 3.

  Neither group had a significant increase in Hb levels after 28 days of treatment. However, as shown in Table 3, mice treated with ActRIIB-Fc had a significant increase in red blood cell distribution width (RDW), reflecting the extent of red blood cell size variation in the sample. Surprisingly, mice treated with anti-GDF8 antibody, anti-activin A antibody, or anti-GDF8 + anti-activin A antibody combination did not show a clear increase in% RDW compared to isotype control treated mice. Thus, these experiments showed that antagonism of GDF8 or activin A alone, or the combination of anti-GDF8 antibody + anti-activin A antibody, did not result in the hematological phenotype observed with ActRIIB-Fc treatment.

  To further investigate the side effect differences between ActRIIB-Fc treated subjects and anti-GDF8 + anti-activin A combination treated subjects, microarray and protein expression studies were performed.

  Microarray analysis was performed on skeletal muscle samples from mice treated with isotype controls, ActRIIB-Fc, and H4H1657N2 (anti-GDF8). From these experiments, a set of genes that are uniquely affected by ActRIIB-Fc were identified. Upregulation of endoglin mRNA levels in skeletal muscle of samples from ActRIIB-Fc treated subjects was particularly interesting. Endoglin is a transmembrane protein expressed primarily in endothelial cells and interacts and promotes signal transduction through receptors of the TGFβ family (ALK1) corresponding to TGFβ, BMP9, or BMP10. Signaling mediated by Alk1 and endoglin in endothelial cells is required to maintain normal vasculature. Mutations in the human Alk1 and endoglin genes cause hereditary hemorrhagic telangiectasia (HHT). Patients with HHT exhibit a vascular phenotype including vasodilation and bleeding of the nasal, oral, and gastrointestinal mucosa. Thus, elevated endoglin levels caused by ActRIIB-Fc may reflect at least some of the adverse side effects observed in patients treated with this therapeutic agent.

  Next, experiments were performed to confirm that changes observed in endoglin mRNA were also reflected in protein expression levels using muscle samples from previous experiments. Quantitative Western blot analysis of endoglin protein levels was performed on samples from mice treated with isotype controls, ActRIIB-Fc, H4H1657N2 (anti-GDF8), MAB3381 (anti-activin A), and H4H1657N2 + MAB3381 combination. To confirm that ActRIIB-Fc treatment did not increase the endothelial compartment, endoglin expression was normalized by the endothelial cell marker CD31. The results (normalized with isotype control values) are summarized in Table 4.

  As shown in Table 4, endoglin protein levels were significantly elevated in the ActRIIB-hFc group but not in the anti-GDF8 or anti-activin A treatment groups. Interestingly, endoglin levels were not elevated even in the anti-GDF8 + anti-activin A combination treatment group.

Summary and Conclusions The results shown in the previous example (Example 1) indicate that the combination of anti-GDF8 + anti-activin A treatment can produce a muscle hypertrophy effect that is at least equivalent to that observed with ActRIIB-Fc treatment. Indicates. This Example 2 shows that no adverse side-effect indicators of ActRIIB-Fc treatment such as increased RDW and increased endoglin expression were observed with anti-GDF8, anti-activin A, or anti-GDF8 + anti-activin A combination treatments. . Thus, the inventors surprisingly found that treatment with a GDF8-specific binding protein, or an activin A-specific binding protein, or a combination of a GDF8-specific binding protein and an activin A-specific binding protein, can be performed using ActRIIB- It has been found to provide a very effective way to increase muscle mass and strength, avoiding the harmful side effects caused by Fc.

Example 3 Effect of GDF8 and Activin A Antagonists on Wound Healing Drugs that function to increase muscle mass and strength, such as GDF8 antagonists and activin A antagonists, have patients undergoing surgery (or will undergo surgery) In the setting, for example, it has utility for joint replacement or repair. As such, agents administered to promote muscle mass rescue would ideally not interfere with other aspects of post-operative recovery such as wound healing.

  Therefore, experiments were performed to determine the effects of GDF8 blockade, activin A blockade, and combinations thereof on wound healing compared to the effects of ActRIIB-Fc treatment. These studies were performed with SCID mice. In particular, the effect of administration of H4H1657N2 (anti-GDF8) and MAB3381 (anti-activin A) on wound healing as a single treatment or in combination with each other of the decoy receptor ActRIIB-hFc (SEQ ID NO: 27), which acts more broadly Compared with wound healing effect. Briefly, a circular skin excision wound was created on the left flank of 30 male SCID mice approximately 7-8 weeks old. The animals were divided into 5 treatment groups (n = 6 per group), each receiving 5 subcutaneous injections of isotype control antibody, ActRIIB-hFc, H4H1657N2, MAB3381, or H4H1657N2 + MAB3381. All reagents were administered at 10 mg / kg every 3-4 days. The first dose was given the day before the animal's wound and the final dose was given 2 days before the end of the study on day 14. Digital images of wounds were taken on days 0 (wound days), 6, 9, 12, and 14 to measure changes in resected wound size and compared to an isotype control group. The results are summarized in Table 5. All data were expressed as mean total wound size ± standard error of the mean.

  As shown in Table 5, as a result of wound size analysis at the end of the experiment, treatment with H4H1657N2, MAB3381, or H4H1657N2 + MAB3381 showed a significant difference in wound size at any time after the initial resection compared to the isotype control group. Did not bring. In contrast, ActRIIB-hFc is wounded as indicated by larger wound sizes at 6, 9, 12, and 14 days compared to wound healing in mice treated with H4H1657N2, MAB3381, H4H1657N2 + MAB3381, or isotype control. Closure was significantly delayed.

  From this experiment, therapeutic treatments including GDF8 antagonism, activin A antagonism, or GDF8 + activin A dual antagonism do not significantly repair wound healing, while the smaller specificity antagonist ActRIIB-hFc Demonstrated significant repair of wound healing. Thus, this example shows that specific antagonism of GDF8 and activin A can lead to increased muscle mass and muscle function similar to that seen with ActRIIB-hFc treatment, but with ActRIIB-hFc treatment Provides further support for the notion of no associated adverse side effects.

  The present invention is not to be limited in scope by the specific embodiments described herein. Indeed, various modifications of the method in addition to the methods described herein will become apparent to those skilled in the art from the foregoing description and accompanying drawings. Such modifications are intended to fall within the scope of the claims.

Claims (20)

A composition for increasing muscle mass or strength, comprising a GDF8 specific binding protein and an activin A specific binding protein comprising:
The GDF8-specific binding protein is an anti-GDF8 antibody or antigen-binding fragment thereof, and the activin A-specific binding protein is an anti-activin A antibody or antigen-binding fragment thereof;
The anti-GDF8 antibody or antigen-binding fragment thereof comprises a heavy chain complementarity determining region (HCDR) of heavy chain variable region (HCVR) comprising SEQ ID NO: 9 and a light chain complement of light chain variable region (LCVR) comprising SEQ ID NO: 13. A composition comprising a sex determining region (LCDR). The HCVR of the anti-GDF8 antibody or antigen-binding fragment thereof comprises the amino acid sequences of HCDR1, HCDR2, and HCDR3 consisting of SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12, respectively, and the LCVR of the anti-GDF8 antibody or antigen-binding fragment thereof The composition for increasing muscle mass or muscle strength according to claim 1, comprising an amino acid sequence of LCDR1, LCDR2, and LCDR3 consisting of SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16, respectively. An antigen binding molecule comprising a GDF8 specific binding domain and an activin A specific binding domain comprising:
The GDF8 specific binding domain includes a heavy chain variable region (HCVR) and a light chain variable region (LCVR), and an activin A specific binding domain includes a heavy chain variable region (HCVR) and a light chain variable region (LCVR). Including
The HCVR of the GDF8 specific binding domain comprises the amino acid sequences of HCDR1, HCDR2, and HCDR3 consisting of SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12, respectively, and the LCVR of the anti-GDF8 specific binding domain is SEQ ID NO: 14, comprising the amino acid sequences of LCDR1, LCDR2, and LCDR3 consisting of SEQ ID NO: 15 and SEQ ID NO: 16,
Antigen binding molecule.   4. The antigen binding molecule of claim 3, wherein the antigen binding molecule is a bispecific antibody. Use of a GDF8 specific binding protein and an activin A specific binding protein in the manufacture of a medicament for increasing a subject's muscle mass or strength comprising:
The GDF8-specific binding protein is an anti-GDF8 antibody or antigen-binding fragment thereof, and the activin A-specific binding protein is an anti-activin A antibody or antigen-binding fragment thereof;
The anti-GDF8 antibody or antigen-binding fragment thereof comprises a heavy chain complementarity determining region (HCDR) of heavy chain variable region (HCVR) comprising SEQ ID NO: 9 and a light chain complement of light chain variable region (LCVR) comprising SEQ ID NO: 13. Including a sex determining region (LCDR),
use.   The anti-GDF8 antibody or antigen-binding fragment thereof comprises HCDR1, HCDR2, and HCDR3 amino acid sequences consisting of SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12, respectively, and SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16, respectively. The use according to claim 5, comprising the amino acid sequences of LCDR1, LCDR2, and LCDR3. Use of an antigen binding molecule comprising a GDF8 specific binding domain and an activin A specific binding domain in the manufacture of a medicament for increasing a subject's muscle mass or strength comprising:
The GDF8 specific binding domain includes a heavy chain variable region (HCVR) and a light chain variable region (LCVR), and an activin A specific binding domain includes a heavy chain variable region (HCVR) and a light chain variable region (LCVR). Including
The HCVR of the GDF8 specific binding domain comprises the amino acid sequences of HCDR1, HCDR2, and HCDR3 consisting of SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12, respectively, and the LCVR of the anti-GDF8 specific binding domain is SEQ ID NO: 14, comprising the amino acid sequences of LCDR1, LCDR2, and LCDR3 consisting of SEQ ID NO: 15 and SEQ ID NO: 16,
use.   8. Use according to claim 7, wherein the antigen binding molecule is a bispecific antibody. Use of a GDF8-specific binding protein and an activin A-specific binding protein in the manufacture of a medicament for treating diseases and disorders characterized by muscle mass or weakness, comprising
The GDF8-specific binding protein is an anti-GDF8 antibody or antigen-binding fragment thereof, and the activin A-specific binding protein is an anti-activin A antibody or antigen-binding fragment thereof;
The anti-GDF8 antibody or antigen-binding fragment thereof comprises a heavy chain complementarity determining region (HCDR) of heavy chain variable region (HCVR) comprising SEQ ID NO: 9 and a light chain complement of light chain variable region (LCVR) comprising SEQ ID NO: 13. Including a sex determining region (LCDR),
use.   Diseases and disorders characterized by muscle mass or weakness include sarcopenia, cachexia, muscle damage, muscle wasting and atrophy, cancer, obesity, diabetes, arthritis, multiple sclerosis, muscular dystrophy, amyotrophic lateral cord 10. Use according to claim 9, selected from the group consisting of sclerosis, Parkinson's disease, osteoporosis, osteoarthritis, osteopenia and metabolic syndrome.   11. Use according to claim 10, wherein the cachexia is idiopathic or secondary to other conditions selected from cancer, chronic renal failure, or chronic obstructive pulmonary disease. Claims wherein muscle wasting and atrophy are caused by or associated with the need for disuse, immobilization, bed rest, injury, medical treatment or surgical intervention, artificial respiration, medical treatment, or surgical intervention. 10. Use according to 10.   13. Use according to claim 12, wherein the medical treatment or surgical intervention is selected from hip fracture, hip replacement, or knee replacement.   11. Use according to claim 10, wherein the metabolic syndrome is selected from the group consisting of diabetes, obesity, nutritional disorders, organ atrophy, chronic obstructive pulmonary disease, and anorexia. Use of an antigen-binding molecule comprising a GDF8-specific binding domain and an activin A-specific binding domain in the manufacture of a medicament for treating diseases and disorders characterized by muscle mass or weakness, comprising
The GDF8 specific binding domain includes a heavy chain variable region (HCVR) and a light chain variable region (LCVR), and an activin A specific binding domain includes a heavy chain variable region (HCVR) and a light chain variable region (LCVR). Including
The HCVR of the GDF8 specific binding domain comprises the amino acid sequences of HCDR1, HCDR2, and HCDR3 consisting of SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12, respectively, and the LCVR of the anti-GDF8 specific binding domain is SEQ ID NO: 14, comprising the amino acid sequences of LCDR1, LCDR2, and LCDR3 consisting of SEQ ID NO: 15 and SEQ ID NO: 16,
use.   Diseases and disorders characterized by muscle mass or weakness include sarcopenia, cachexia, muscle damage, muscle wasting and atrophy, cancer, obesity, diabetes, arthritis, multiple sclerosis, muscular dystrophy, amyotrophic lateral cord 16. Use according to claim 15, selected from the group consisting of sclerosis, Parkinson's disease, osteoporosis, osteoarthritis, osteopenia and metabolic syndrome.   16. Use according to claim 15, wherein the cachexia is idiopathic or secondary to other conditions selected from cancer, chronic renal failure, or chronic obstructive pulmonary disease.   Claims wherein muscle wasting and atrophy are caused by or associated with the need for disuse, immobilization, bed rest, injury, medical treatment or surgical intervention, artificial respiration, medical treatment, or surgical intervention. 15. Use according to 15.   19. Use according to claim 18, wherein the medical treatment or surgical intervention is selected from hip fracture, hip replacement, or knee replacement.   16. The use according to claim 15, wherein the metabolic syndrome is selected from the group consisting of diabetes, obesity, malnutrition, organ atrophy, chronic obstructive pulmonary disease, and anorexia.